Cloning and Expression Analysis of As6G-FFT Gene Encoding Fructan Synthase from Garlic

doi:10.16420/j.issn.0513-353x.2021-0403

Acta Horticulturae Sinica ›› 2021, Vol. 48 ›› Issue (12): 2403-2413.doi: 10.16420/j.issn.0513-353x.2021-0403

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Cloning and Expression Analysis of As6G-FFT Gene Encoding Fructan Synthase from Garlic

TIAN Jie¹^,²(), BIAN Haiyan¹, TIE Yuanyu¹

¹Qinghai Key Laboratory of Vegetable Genetics and Physiology,Qinghai Academy of Agricultural and Forestry Sciences,Xining 810016,China
²State Key Laboratory of Plateau Ecology and Agriculture,Xining 810016,China

Received:2021-07-26 Revised:2021-12-03 Published:2022-01-04

Abstract

Abstract:

Fructan is the main storage material of garlic,and its metabolism is closely related to yield formation and stress adaptability. In order to understand the sequence characteristics and expression patterns of fructan：fructan 6G-Fructosyltransferase gene（As6G-FFT）in different tissues and stress conditions of garlic,the As6G-FFT（fructan：fructan 6G-Fructosyltransferase）gene was cloned from ‘Ledu Purple Skin Garlic’. The CDS full length of As6G-FFT gene was 1 839 bp,encoding 612 amino acids with molecular mass of 68.64 and theoretical isoelectric point of 5.43. It was a hydrophilic protein and contained three important conserved domains of NDPSG,RDP and EC of plant glycosylhydrolase 32 family. In terms of evolutionary relationship,garlic As6G-FFT gene was close to onion. The result of subcellular localization showed that As6G-FFT protein was located in endoplasmic reticulum. qRT-PCR analysis showed that As6G-FFT was expression in all tissues examined,which expression level was leaf > root > mother petal > pseudostem. In addition,the expression characteristics of As6G-FFT gene in different tissues were also different after low temperature（4 ℃）or drought stress treatment（45%-55% relative water content of matrix）. Compared with other tissues,drought stress could induce the expression of As6G-FFT gene in roots earlier,whereas the expression of As6G-FFT in mother petal was more sensitive under low temperature stress,which indicated that the response mechanism of As6G-FFT in garlic tissues to stress signals was different.

Key words: garlic, fructan, As6G-FFT gene, gene cloning, expression analysis

CLC Number:

S633.4

TIAN Jie, BIAN Haiyan, TIE Yuanyu. Cloning and Expression Analysis of As6G-FFT Gene Encoding Fructan Synthase from Garlic[J]. Acta Horticulturae Sinica, 2021, 48(12): 2403-2413.

Figures/Tables 8

Fig. 1 PCR amplification of As6G-FFT

Fig. 2 Nucleotide sequences and predicted amino acid sequences of As6G-FFT from garlic

Fig. 3 The contents of amino acids of As6G-FFT protein sequence from garlic

Fig. 4 Analysis of 6G-FFT conserved motifs in garlic and other plants

Fig. 5 Comparative analysis of 6G-FFT amino acid sequences between garlic and other plants The red box indicates three conserved motifs：NDPNG,RDP and EC.

Fig. 6 Subcellular localization analysis of Allium sativum As6G-FFT protein

Fig. 7 Relative expression level of As6G-FFT gene in different garlic tissues P < 0.05.。。

Fig. 8 Relative expression level of as6g FFT gene in different garlic tissues under low temperature and drought stress P < 0.05.

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Cloning and Expression Analysis of As6G-FFT Gene Encoding Fructan Synthase from Garlic

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